Methods for isolating and analyzing physiological hyaluronan: a review

Rivas, Felipe; Erxleben, Dorothea; Smith, Ian; Rahbar, Elaheh; DeAngelis, Paul J.; Cowman, Mary K.; Hall, Adam R.

doi:10.1152/ajpcell.00019.2022

Cited by 9 publications

(10 citation statements)

References 192 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Gas-phase electrophoretic mobility molecular analysis (GEMMA) provides mean size and size dispersity, and requires a much smaller amount of sample (pg range) 15 . Gel electrophoresis (GE) can be performed in any biochemistry lab and can be adapted to virtually any GAG size (with sample needs in the upper ng to lower μg range, dependent on size distribution and staining method), but requires a set of size standards for analysis 14 , 16 , 17 . Importantly, GEMMA and GE are strongly affected by GAG charge in addition to size which limits the use of these methods for sulfated GAG polysaccharides with varying charge distribution.…”

Section: Introductionmentioning

confidence: 99%

A quartz crystal microbalance method to quantify the size of hyaluronan and other glycosaminoglycans on surfaces

Srimasorn

Souter

Green

et al. 2022

Sci Rep

View full text Add to dashboard Cite

Hyaluronan (HA) is a major component of peri- and extra-cellular matrices and plays important roles in many biological processes such as cell adhesion, proliferation and migration. The abundance, size distribution and presentation of HA dictate its biological effects and are also useful indicators of pathologies and disease progression. Methods to assess the molecular mass of free-floating HA and other glycosaminoglycans (GAGs) are well established. In many biological and technological settings, however, GAGs are displayed on surfaces, and methods to obtain the size of surface-attached GAGs are lacking. Here, we present a method to size HA that is end-attached to surfaces. The method is based on the quartz crystal microbalance with dissipation monitoring (QCM-D) and exploits that the softness and thickness of films of grafted HA increase with HA size. These two quantities are sensitively reflected by the ratio of the dissipation shift (ΔD) and the negative frequency shift (− Δf) measured by QCM-D upon the formation of HA films. Using a series of size-defined HA preparations, ranging in size from ~ 2 kDa tetrasaccharides to ~ 1 MDa polysaccharides, we establish a monotonic yet non-linear standard curve of the ΔD/ − Δf ratio as a function of HA size, which reflects the distinct conformations adopted by grafted HA chains depending on their size and surface coverage. We demonstrate that the standard curve can be used to determine the mean size of HA, as well as other GAGs, such as chondroitin sulfate and heparan sulfate, of preparations of previously unknown size in the range from 1 to 500 kDa, with a resolution of better than 10%. For polydisperse samples, our analysis shows that the process of surface-grafting preferentially selects smaller GAG chains, and thus reduces the average size of GAGs that are immobilised on surfaces comparative to the original solution sample. Our results establish a quantitative method to size HA and other GAGs grafted on surfaces, and also highlight the importance of sizing GAGs directly on surfaces. The method should be useful for the development and quality control of GAG-based surface coatings in a wide range of research areas, from molecular interaction analysis to biomaterials coatings.

show abstract

Section: Introductionmentioning

confidence: 99%

A quartz crystal microbalance method to quantify the size of hyaluronan and other glycosaminoglycans on surfaces

Srimasorn

Souter

Green

et al. 2022

Sci Rep

View full text Add to dashboard Cite

show abstract

“…Several methods are now well established to quantify the mass of HA and other GAGs in the solution phase [14]. Multi-angle light scattering (MALS) when coupled with size exclusion chromatography (SEC) or field flow fractionation (FFF) provides weight-averaged ( M W ) and number-averaged ( M n ) molecular masses, and thus also the polydispersity index (PDI = M W / M n ) of a sample without the need for a size standard but requires relatively large amounts of sample (typically tens to hundreds of μg for SEC-MALS [14]). Gas-phase electrophoretic mobility molecular analysis (GEMMA) provides mean size and size dispersity, and requires a much smaller amount of sample (pg range) [15].…”

Section: Introductionmentioning

confidence: 99%

“…Gas-phase electrophoretic mobility molecular analysis (GEMMA) provides mean size and size dispersity, and requires a much smaller amount of sample (pg range) [15]. Gel electrophoresis (GE) can be performed in any biochemistry lab and can be adapted to virtually any GAG size (with sample needs in the upper ng to lower μg range, dependent on size distribution and staining method), but requires a set of size standards for analysis [14, 16, 17]. Importantly, GEMMA and GE are strongly affected by GAG charge in addition to size which limits the use of these methods for sulphated GAG polysaccharides with varying charge distribution.…”

Section: Introductionmentioning

confidence: 99%

“…In light of the wide range of GAG sizes and their biological importance, the ability to quantify the size of GAGs is key to progress in GAG biology and the use of GAGs in medical, pharmaceutical and cosmetic applications. Several methods are now well established to quantify the mass of HA and other GAGs in the solution phase [14]. Multi-angle light scattering (MALS) when coupled with size exclusion chromatography (SEC) or field flow fractionation (FFF) provides weight-averaged ( M W ) and number-averaged ( M n ) molecular masses, and thus also the polydispersity index (PDI = M W / M n ) of a sample without the need for a size standard but requires relatively large amounts of sample (typically tens to hundreds of μg for SEC-MALS [14]).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

A quartz crystal microbalance method to quantify the size of hyaluronan and other glycosaminoglycans on surfaces

Srimasorn

Souter

Green

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

Hyaluronan (HA) is a major component of peri- and extra-cellular matrices and plays important roles in many biological processes such as cell adhesion, proliferation and migration. The abundance, size distribution and presentation of HA dictate its biological effects and are also useful indicators of pathologies and disease progression. Methods to assess the molecular mass of soluble HA and other glycosaminoglycans (GAGs) are well established. In many biological and technological settings, however, GAGs are displayed on surfaces, and methods to obtain the size of surface-attached GAGs are lacking. Here, we present a method to size HA that is end-attached to surfaces. The method is based on the quartz crystal microbalance with dissipation monitoring (QCM-D) and exploits that the softness and thickness of films of grafted HA increase with HA size. These two quantities are sensitively reflected by the ratio of the dissipation shift (ΔD) and the negative frequency shift (-Δf) measured by QCM-D upon the formation of HA films. Using a series of size-defined HA preparations, ranging in size from a tetrasaccharide to a 1 MDa polysaccharide, we establish a standard curve of the ΔD/-Δf ratio as a function of HA size. We demonstrate that the standard curve can be used to determine the mean size of HA, as well as other GAGs, such as chondroitin sulfate and heparan sulfate, of preparations of previously unknown size in the range from 1 to 500 kDa, and with a resolution better than 10%. For polydisperse samples, our analysis shows that the process of surface-grafting preferentially selects smaller GAG chains, and thus reduced the average size of GAGs that are immobilised on surfaces comparative to the original solution sample. Our results establish a quantitative method to size HA and other GAGs grafted on surfaces, and also highlight the importance of sizing GAGs directly on surfaces. The method should be useful for the development and quality control of GAG-based surface coatings in a wide range of research areas, from molecular interaction analysis to biomaterials coatings.

show abstract

“…Hyaluronic acid (HA) is a nonsulfated linear glycosaminoglycan (GAG) with a negative charge and comprising a tandem repeating unit of the disaccharide β-1,4-D-glucuronic acid-β-1,3-D-N-acetylglucosamine-β-1,4. 1,2 HA is abundant in the extracellular matrix of cells, with the most distribution around skin cells, connective tissue, joint tissue, and vitreous body. 3,4 A distinct difference between the high-molecular-weight HA polymers (HMW-HAs, ∼2 × 10 3 kDa) and the low-molecularweight HAs (LMW-HAs, 4−120 kDa) and HA oligosaccharides (O-HAs, <4 kDa) is the unique biological properties exhibited by the latter two.…”

Section: Introductionmentioning

confidence: 99%

Enzymatic Production of Low-Molecular-Weight Hyaluronan and Its Oligosaccharides: A Review and Prospects

Pang

Wang²,

Huang

et al. 2022

J. Agric. Food Chem.

View full text Add to dashboard Cite

Hyaluronic acid (HA) is a nonsulfated linear glycosaminoglycan with a negative charge. Different from the high-molecular-weight HAs, the low-molecular-weight HAs (LMW-HAs, 4–120 kDa) and hyaluronan oligosaccharides (O-HAs, <4 kDa) exhibit certain unique biological properties, owing to which these have a wide range of applications in the field of medicine. However, the chemical synthesis of high-purity LMW-HAs and O-HAs requires complex procedures, which renders this process difficult to achieve. The degradation of HA is achieved under the catalysis of hyaluronidases. In recent years, various hyaluronidase genes have been identified, and their enzymatic properties have been analyzed. In this context, the present review summarizes the hyaluronidases from different sources, which have been characterized. The review focuses on the crystal structure and the catalytic mechanism underlying the biological properties of hyaluronidases. In addition, the molecular weight distributions and the preparation approaches of the enzymatic products LMW-HAs and O-HAs are described. The general orientation of the research on hyaluronidases was speculated based on the existing literature. Accordingly, the efficient large-scale production of LMW-HAs and O-HAs using the green enzymatic approach was anticipated.

show abstract

Methods for isolating and analyzing physiological hyaluronan: a review

Cited by 9 publications

References 192 publications

A quartz crystal microbalance method to quantify the size of hyaluronan and other glycosaminoglycans on surfaces

A quartz crystal microbalance method to quantify the size of hyaluronan and other glycosaminoglycans on surfaces

A quartz crystal microbalance method to quantify the size of hyaluronan and other glycosaminoglycans on surfaces

Enzymatic Production of Low-Molecular-Weight Hyaluronan and Its Oligosaccharides: A Review and Prospects

Contact Info

Product

Resources

About